LOCUS       sequence-388914-       13491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NRCH-ABE8e(V106W) and 
            blasticidin resistance: EFS 
            promoter-SpCas9-NRCH-ABE8e(V106W)-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13491)
  AUTHORS   Kim N, Choi S, Kim S, Song M, Seo JH, Min S, Park J, Cho SR, Kim HH
  TITLE     Deep learning models to predict the editing efficiencies and 
            outcomes of diverse base editors.
  JOURNAL   Nat Biotechnol. 2024 Mar;42(3):484-497. doi: 
            10.1038/s41587-023-01792-x
  PUBMED    37188916
REFERENCE   2  (bases 1 to 13491)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nat Biotechnol"; date: "2024-03"; volume: "42"; issue: "3"; pages: 
            "484-497"
COMMENT     Sequence Label: SpCas9-NRCH-ABE8e(V106W)
FEATURES             Location/Qualifiers
     source          1..13491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        55..434
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        436..634
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     589..609
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             652..832
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    879..1004
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1497..1730
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1915..1959
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2257..2374
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2467..2678
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2729..2749
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3344..7444
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMVKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQGDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRLRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGGH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKGNSDKLIARKKDWDPKKYGGFNSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTINRKQYNTTKEVLD
                     ATLIRQSITGLYETRIDLSQLGGD"
     CDS             7487..7507
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7508..7531
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7541..7597
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7598..7996
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8021..8609
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8074..8094)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8492..8503)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8612..8628)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8613..8629)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9134..9314
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9340..9357)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9346..9570
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9616..10044
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9703..9722)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9913..9934
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10053..10073)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      10218..10352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10279..10298
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10414..10461
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10480..10854
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    10984..11105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11021..11040)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11075..11094
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11154..11170)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11154..11170)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11167..11189)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11178..11194
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(11202..11208,11209..11226,11227..11232))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11247..11268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11385..11402)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11556..12144)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11636..11655)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12315..13106,13107..13175))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     12938..12957
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13176..13280)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13355..13374)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
       61 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      121 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      181 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      241 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      301 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      361 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      421 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      481 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      541 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      601 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
      661 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      721 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      781 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      841 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      901 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      961 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     1021 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     1081 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     1141 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     1201 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     1261 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     1321 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
     1381 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1441 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1501 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1561 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1621 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1681 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1741 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1801 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1861 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1921 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1981 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     2041 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     2101 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     2161 ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg
     2221 ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     2281 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     2341 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     2401 tccagtttgg ttaattagct agctaggtct tgaaaggagt gggaattggc tccggtgccc
     2461 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     2521 attgatccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     2581 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     2641 acgttctttt tcgcaacggg tttgccgcca gaacacagga ccggttgcca ccatgaaacg
     2701 gacagccgac ggaagcgagt tcgagtcacc aaagaagaag cggaaagtct ctgaggtgga
     2761 gttttcccac gagtactgga tgagacatgc cctgaccctg gccaagaggg cacgggatga
     2821 gagggaggtg cctgtgggag ccgtgctggt gctgaacaat agagtgatcg gcgagggctg
     2881 gaacagagcc atcggcctgc acgacccaac agcccatgcc gaaattatgg ccctgagaca
     2941 gggcggcctg gtcatgcaga actacagact gattgacgcc accctgtacg tgacattcga
     3001 gccttgcgtg atgtgcgccg gcgccatgat ccactctagg atcggccgcg tggtgtttgg
     3061 atggagaaat tctaaaagag gcgccgcagg ctccctgatg aacgtgctga actaccccgg
     3121 catgaatcac cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg ccgccctgct
     3181 gtgcgatttc tatcggatgc ctagacaggt gttcaatgct cagaagaagg cccagagctc
     3241 catcaactcc ggaggatcta gcggaggctc ctctggctct gagacacctg gcacaagcga
     3301 gagcgcaaca cctgaaagca gcgggggcag cagcgggggg tcagacaaga agtacagcat
     3361 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
     3421 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
     3481 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
     3541 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
     3601 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
     3661 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
     3721 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
     3781 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
     3841 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
     3901 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
     3961 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
     4021 ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg
     4081 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
     4141 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
     4201 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
     4261 agtgaacacc gagatcacca aggcccccct gagcgcctct atggtgaaga gatacgacga
     4321 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
     4381 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
     4441 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
     4501 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
     4561 cggcatcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcaggg
     4621 cgatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
     4681 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
     4741 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
     4801 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
     4861 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
     4921 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
     4981 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
     5041 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
     5101 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
     5161 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
     5221 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
     5281 gatgaagcag ctgaagcggc tgagatacac cggctggggc aggctgagcc ggaagctgat
     5341 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
     5401 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
     5461 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
     5521 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
     5581 cgtgaaagtg atgggcggcc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
     5641 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
     5701 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
     5761 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
     5821 actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct
     5881 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag
     5941 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
     6001 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
     6061 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
     6121 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
     6181 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
     6241 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
     6301 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
     6361 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
     6421 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
     6481 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
     6541 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
     6601 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
     6661 agagtctatc ctgcccaagg gcaacagcga taagctgatc gccagaaaga aggactggga
     6721 ccctaagaag tacggcggct tcaacagccc caccgtggcc tattctgtgc tggtggtggc
     6781 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctac tggggatcac
     6841 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
     6901 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
     6961 aaacggccgg aagagaatgc tggcctctgc cggcgtgctg cagaagggaa acgaactggc
     7021 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
     7081 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
     7141 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
     7201 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
     7261 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
     7321 tgacaccacc atcaaccgga agcagtacaa caccaccaaa gaggtgctgg acgccaccct
     7381 gatccggcag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
     7441 cgactctggc ggctcaaaaa gaaccgccga cggcagcgaa ttcgagccca agaagaagag
     7501 gaaagtcgat tacaaagacg atgacgataa gggatccggc gcaacaaact tctctctgct
     7561 gaaacaagcc ggagatgtcg aagagaatcc tggaccgatg gccaagcctt tgtctcaaga
     7621 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
     7681 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
     7741 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
     7801 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
     7861 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
     7921 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
     7981 tgtgtgggag ggctaagaat tcgatatcaa gcttatcggt aatcaacctc tggattacaa
     8041 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     8101 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     8161 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     8221 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     8281 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     8341 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     8401 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     8461 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     8521 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     8581 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag acctagaaaa
     8641 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
     8701 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
     8761 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     8821 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
     8881 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt
     8941 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg
     9001 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     9061 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca
     9121 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     9181 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     9241 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     9301 gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     9361 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     9421 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     9481 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     9541 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     9601 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     9661 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     9721 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     9781 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     9841 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     9901 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     9961 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
    10021 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
    10081 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
    10141 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
    10201 ttagtcagca accatagtcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
    10261 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
    10321 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
    10381 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
    10441 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc
    10501 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
    10561 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
    10621 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
    10681 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
    10741 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
    10801 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg
    10861 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
    10921 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
    10981 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
    11041 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
    11101 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
    11161 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
    11221 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
    11281 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
    11341 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
    11401 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
    11461 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
    11521 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
    11581 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
    11641 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
    11701 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
    11761 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
    11821 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
    11881 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
    11941 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
    12001 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
    12061 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
    12121 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
    12181 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
    12241 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
    12301 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
    12361 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
    12421 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
    12481 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
    12541 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
    12601 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
    12661 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
    12721 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
    12781 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
    12841 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
    12901 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
    12961 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
    13021 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
    13081 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
    13141 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
    13201 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
    13261 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
    13321 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
    13381 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
    13441 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg c
//
